Prothrombin, alone or in complex concentrates or plasma, reduces bleeding in a mouse model of blood exchange-induced coagulopathy

Abstract

Prothrombin complex concentrates (PCC) are fractionated plasma protein drugs that reverse warfarin anticoagulation. PCC may control more general bleeding. We sought to identify the dominant procoagulant factor in PCC in vivo. We tested PCC or coagulation factor (F) treatment in CD1 mice made coagulopathic by exchange of whole blood for washed red cells. Anesthetized mice were transfused with murine fresh-frozen plasma (mFFP), PCC, mixtures of human vitamin K-dependent proteins (VKDP) (prothrombin, FVII, FIX, or FX), or purified single human VKDP, immediately prior to tail transection (TT), liver laceration (LL), or intravascular laser injury (ILI). Plasma donor mice were treated with vehicle or control antisense oligonucleotide (ASO-CON) or ASO specific for prothrombin (FII) (ASO-FII) to yield mFFP or ASO-CON mFFP or ASO-FII mFFP. Blood losses were determined spectrophotometrically (TT) or gravimetrically (LL). Thrombus formation was quantified by intravital microscopy of laser-injured arterioles. PCC or four factor- (4F-) VKDP or prothrombin significantly reduced bleeding from TT or LL. Omission of prothrombin from 4F-VKDP significantly reduced its ability to limit bleeding. Mice transfused with ASO-FII mFFP demonstrated inferior haemostasis versus those transfused with ASO-FII following TT, LL, or ILI. Prothrombin is the dominant procoagulant component of PCC and could limit bleeding in trauma.

The association of thrombin generation with bleeding outcomes in cardiac surgery: a prospective observational study

“…35 While further research is needed, the optimal treatment of decreased thrombin generation in cardiac surgical patients may eventually include low-dose coagulation factor isolates. 36 Our study suggests the ETP in particular may be a useful therapeutic target.…”

Correction of haemorrhagic shock-associated coagulopathy and impaired haemostasis by plasma, prothrombin complex concentrates or an activated protein C-targeted DNA aptamer in mice

“…PCC therefore likely exerts a procoagulant effect by increasing the concentrations of the procoagulant factors listed above, facilitating their activation by mass action. We previously found PCC to reduce bleeding and eliminate coagulopathy in a normovolemic mouse model of pan-plasma factor depletion achieved by sequential blood exchange 23 .…”

“…Because of shock-related peripheral vasoconstriction, we could not assess haemostasis via tail transection, finding in initial studies that HS/LL resuscitated mice bled very little from this peripheral injury. We therefore selected standardized liver laceration as the haemostatic challenge, based on our previous work in mice rendered coagulopathic via anticoagulant administration 43 or blood exchange 22,23 .…”

“…Sample sizes. Sample sizes were determined via power calculations (GraphPad Statmate 2.0, GraphPad Software, San Diego, CA, USA), using pilot data from our previous model of murine coagulopathy and liver laceration 23 , which revealed that with α at 0.05, SD (σ) of 33.5 mg, n = 6 was predicted to give 80% power to detect a significant difference in means of 65 mg of blood lost.…”